家畜诱导性多能干细胞培养体系研究进展

doi:10.19543/j.cnki.1673-4556.20260203.001

摘要/Abstract

摘要：

诱导性多能干细胞（Induced pluripotent stem cells，iPSCs）的建立为干细胞的研究带来了革命性突破。经过多年发展，相比于人和小鼠iPSCs成熟的重编程技术与培养方法，家畜iPSCs虽可在体外维持增殖并具有限定分化能力，但尚未突破生殖嵌合等体内多能性金标准，且培养体系、标志物明确性及分化效率等均显著落后。该文介绍了家畜iPSCs重编程过程中的起始材料与方法，阐述了各类成体细胞在家畜iPSCs诱导中的应用与效率差异，分析了不同载体的优劣以及重编程因子在安全性与效率方面的作用，并详细梳理了饲养层细胞、基础培养基、小分子化合物和培养环境等培养条件对家畜iPSCs稳定性的影响，最后探讨了家畜iPSCs在育种、生物医学及细胞培养肉等方面的应用前景，以期为家畜iPSCs的进一步发展提供思路。

关键词: 诱导性多能干细胞, 重编程因子, 小分子化合物

Abstract:

The establishment of induced pluripotent stem cells (iPSCs) represents a revolutionary breakthrough in stem cell research. After years of development, compared to the mature reprogramming techniques and culture methods for human and murine iPSCs, livestock iPSCs – while capable of sustaining proliferation in vitro and possessing limited differentiation potential – have yet to achieve in vivo pluripotency gold standards such as germline transmission. Furthermore, critical aspects including culture systems, definitive marker characterization, and differentiation efficiency remain significantly underdeveloped. This review outlines the initial materials and methods used in livestock iPSC reprogramming, discusses the applications and efficiency variations of different somatic cell types in generating livestock iPSCs, and evaluates the advantages and limitations of various delivery vectors as well as the roles of reprogramming factors in safety and efficiency. It also systematically examines how culture conditions: including feeder cells, basal media, small molecule compounds, and environmental factors—affect the stability of livestock iPSCs. Finally, the potential applications of livestock iPSCs in breeding, biomedical research, and cell-cultured meat production are explored, providing insights and directions for future advancements in this field.

Key words: Induced pluripotent stem cells, Reprogramming factor, Small molecule compounds

中图分类号:

S813.3

李姝慧, 赵文轩, 高晨, 高雪. 家畜诱导性多能干细胞培养体系研究进展[J]. 中国畜禽种业, 2026, 22(9): 37-46.

Shuhui Li, Wenxuan Zhao, Chen Gao, Xue Gao. Research progress on culture systems of induced pluripotent stem cells in livestock[J]. Chinese Livestock and Poultry Breeding, 2026, 22(9): 37-46.

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参考文献 103

[1]	TAKAHASHI K, YAMANAKA S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J]. Cell, 2006, 126(4): 663-676.
[2]	TAKAHASHI K, TANABE K, OHNUKI M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors[J]. Cell, 2007, 131(5): 861-872.
[3]	GUAN J Y, WANG G, WANG J L, et al. Chemical repro-gramming of human somatic cells to pluripotent stem cells[J]. Nature, 2022, 605(7909): 325-331.
[4]	CHENG Y S, XU M, CHEN G B, et al. A protocol for culture and characterization of human induced pluripotent stem cells after induction[J]. Current Protocols, 2023, 3(8): e866.
[5]	CUESTA-GOMEZ N, VERHOEFF K, DADHEECH N, et al. Suspension culture improves iPSC expansion and pluripotency phenotype[J]. Stem Cell Research and Therapy, 2023, 14(1): 154.
[6]	EZASHI T, TELUGU B P V L, ALEXENKO A P, et al. Derivation of induced pluripotent stem cells from pig somatic cells[J]. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(27): 10993-10998.
[7]	WU Z, CHEN J J, REN J T, et al. Generation of pig induced pluripotent stem cells with a drug-inducible system[J]. Journal of Molecular Cell Biology, 2009, 1(1): 46-54.
[8]	ESTEBAN M A, XU J Y, YANG J Y, et al. Generation of induced pluripotent stem cell lines from Tibetan miniature pig[J]. The Journal of Biological Chemistry, 2009, 284(26): 17634-17640.
[9]	LI Y, CANG M, LEE A S, et al. Reprogramming of sheep fibroblasts into pluripotency under a drug-inducible expression of mouse-derived defined factors[J]. PLoS One, 2011, 6(1): e15947.
[10]	BAO L, HE L, CHEN J J, et al. Reprogramming of ovine adult fibroblasts to pluripotency via drug-inducible expression of defined factors[J]. Cell Research, 2011, 21(4): 600-608.
[11]	SUMER H, LIU J, MALAVER-ORTEGA L F, et al. NANOG is a key factor for induction of pluripotency in bovine adult fibroblasts[J]. Journal of Animal Science, 2011, 89(9): 2708-2716.
[12]	PESSÔA L V F, BRESSAN F F, FREUDE K K. Induced pluripotent stem cells throughout the animal Kingdom: Availability and applications[J]. World Journal of Stem Cells, 2019, 11(8): 491-505.
[13]	RECCHIA K, PESSÔA L V F, PIERI N C G, et al. Influence of cell type in in vitro induced reprogramming in cattle[J]. Life, 2022, 12(8): 1139.
[14]	XIAO L Z, HUANG Z F, WU Z X, et al. Reconstitution of pluripotency from mouse fibroblast through Sall4 overexpression[J]. Nature Communications, 2024, 15: 10787.
[15]	WANG X T, LIU X Y, QU M Y, et al. Sertoli cell-only syndrome: Advances, challenges, and perspectives in genetics and mechanisms[J]. Cellular and Molecular Life Sciences, 2023, 80(3): 67.
[16]	SHI X Y, ZHANG K, YU F S, et al. Advancements and innovative strategies in induced pluripotent stem cell-derived mesenchymal stem cell therapy: A comprehensive review[J]. Stem Cells International, 2024, 2024(1): 4073485.
[17]	JIAMVORAPHONG N, NIMSAMER P, WONGSURAWAT T, et al. Establishment of the integration-free Class I and Class II 11 loci homozygous HLA iPSC line (MUSIi023-A) from peripheral blood mononuclear cells of a Thai donor[J]. Stem Cell Research, 2025, 88: 103840.
[18]	KLINGENSTEIN S, KLEGER A, LIEBAU S, et al. State-of-the-art: Somatic cell sources used for the generation of human induced pluripotent stem cells[J]. Stem Cells and Development, 2025, 34(15/16): 317-332.
[19]	KUMAR A, FISCHER C, CICHOCKI F, et al. Multiplexed iPSC platform for advanced NK cell immunotherapies[J]. Cell Reports Medicine, 2025, 6(11): 102282.
[20]	RECCHIA K, MACHADO L S, BOTIGELLI R C, et al. In vitro induced pluripotency from urine-derived cells in porcine[J]. World Journal of Stem Cells, 2022, 14(3): 231-244.
[21]	KAWAGUCHI T, TSUKIYAMA T, KIMURA K, et al. Generation of Naïve bovine induced pluripotent stem cells using PiggyBac transposition of doxycycline-inducible transcription factors[J]. PLoS One, 2015, 10(8): e0135403.
[22]	CRAVERO D, MARTIGNANI E, MIRETTI S, et al. Generation of induced pluripotent stem cells from bovine epithelial cells and partial redirection toward a mammary phenotype in vitro [J]. Cellular Reprogramming, 2015, 17(3): 211-220.
[23]	ZHAO L X, WANG Z X, ZHANG J D, et al. Characterization of the single-cell derived bovine induced pluripotent stem cells[J]. Tissue & Cell, 2017, 49(5): 521-527.
[24]	HANNA M, SAHITO R G A, RATEB M, et al. Generation of transgene-free induced pluripotent stem cells from cardiac fibroblasts of goat embryos[J]. Journal of Stem Cells & Regene-Rative Medicine, 2020, 16(2): 34-43.
[25]	LIU F, WANG J, YUE Y L, et al. Derivation of arbas Cashmere goat induced pluripotent stem cells in LCDM with trophectoderm lineage differentiation and interspecies chimeric abilities[J]. International Journal of Molecular Sciences, 2023, 24(19): 14728.
[26]	SONG H, LI H, HUANG M R, et al. Induced pluripotent stem cells from goat fibroblasts[J]. Molecular Reproduction and development, 2013, 80(12): 1009-1017.
[27]	JIANG Y, AN X L, YU H, et al. Transcriptome profile of bovine iPSCs derived from Sertoli Cells[J]. Theriogenology, 2020, 146: 120-132.
[28]	MAO J, LIU L. Generation of iPS cells from granulosa cells[J]. Methods in Molecular Biology, 2016, 1357: 451-464.
[29]	YU S, ZHU Z S, SHEN Q Y, et al. Comparative analysis of porcine iPSCs derived from Sertoli cells and fibroblasts[J]. Journal of Cellular Physiology, 2022, 237(12): 4531-4543.
[30]	WEI C, LI X, ZHANG P F, et al. Characterization of porcine partially reprogrammed iPSCs from adipose-derived stem cells[J]. Reproduction, 2015, 149(5): 485-496.
[31]	SU Y, WANG L, FAN Z Q, et al. Establishment of bovine-induced pluripotent stem cells[J].International Journal of Molecular Sciences, 2021, 22(19): 10489.
[32]	REN X, XU J C, XUE Q S, et al. BRG1 enhances porcine iPSC pluripotency through WNT/β-catenin and autophagy pathways[J]. Theriogenology, 2024, 215: 10-23.
[33]	SHEN Q Y, WU X J, CHEN Z Y, et al. TBX3 orchestrates H3K4 trimethylation for porcine induced pluripotent stem cells to totipotent-like stem cells[J]. Journal of Integrative Agriculture, 2025, 24(10): 3978-3996.
[34]	MA K Y, SONG G Q, AN X L, et al. miRNAs promote generation of porcine-induced pluripotent stem cells[J]. Molecular and Cellular Biochemistry, 2014, 389(1): 209-218.
[35]	GAO Y, GUO Y J, DUAN A Q, et al. Optimization of culture conditions for maintaining porcine induced pluripotent stem cells[J]. DNA and Cell Biology, 2014, 33(1): 1-11.
[36]	KUES W A, HERRMANN D, BARG-KUES B, et al. Derivation and characterization of sleeping beauty transposon-mediated porcine induced pluripotent stem cells[J]. Stem Cells and Development, 2013, 22(1): 124-135.
[37]	CONRAD J V, MEYER S, RAMESH P S, et al. Efficient derivation of transgene-free porcine induced pluripotent stem cells enables in vitro modeling of species-specific developmental timing[J]. Stem Cell Reports, 2023, 18(12): 2328-2343.
[38]	JIAO H L, LEE M S, SIVAPATHAM A, et al. Epigenetic regulation of BAF60A determines efficiency of miniature swine iPSC generation[J]. Scientific Reports, 2022, 12: 9039.
[39]	CANIZO J R, VAZQUEZ ECHEGARAY C, KLISCH D, et al. Exogenous human OKSM factors maintain pluripotency gene expression of bovine and porcine iPS-like cells obtained with STEMCCA delivery system[J]. BMC Research Notes, 2018, 11(1): 509.
[40]	CAO H G, YANG P, PU Y, et al. Characterization of bovine induced pluripotent stem cells by lentiviral transduction of reprogramming factor fusion proteins[J]. International Journal of Biological Sciences, 2012, 8(4): 498-511.
[41]	BAI C Y, LI X C, GAO Y H, et al. Melatonin improves reprogramming efficiency and proliferation of bovine-induced pluripotent stem cells[J]. Journal of Pineal Research, 2016, 61(2): 154-167.
[42]	TALLURI T R, KUMAR D, GLAGE S, et al. Derivation and characterization of bovine induced pluripotent stem cells by transposon-mediated reprogramming[J]. Cellular Reprogramm-ing, 2015, 17(2): 131-140.
[43]	ZHANG Y F, HE Y H, WU P, et al. miR-200c-141 enhances sheep kidney cell reprogramming into pluripotent cells by targeting ZEB1[J]. International Journal of Stem Cells, 2021, 14(4): 423-433.
[44]	CHEN H, ZUO Q S, WANG Y J, et al. Inducing goat pluripotent stem cells with four transcription factor mRNAs that activate endogenous promoters[J]. BMC Biotechnology, 2017, 17(1): 11.
[45]	STADTFELD M, NAGAYA M, UTIKAL J, et al. Induced pluripotent stem cells generated without viral integration[J]. Science, 2008, 322(5903): 945-949.
[46]	SUN Y X, SU J, WANG X C, et al. Patient-specific iPSC-derived cardiomyocytes reveal variable phenotypic severity of Brugada syndrome[J]. eBioMedicine, 2023, 95: 104741.
[47]	滕亚迪, 李树静, 余文莉, 等. 基因组无外源整合家畜诱导多能干细胞的生产及应用前景[J]. 农业生物技术学报, 2025, 33(11): 2472-2482.
	TENG Y D, LI S J, YU W L, et al. Generation and application prospects of genome integration-free livestock induced pluripotent stem cells[J]. Journal of Agricultural Biotechnology, 2025, 33(11): 2472-2482.
[48]	LEE Y, JEONG D, HAM S, et al. Generation of integration-free human induced pluripotent stem cells from a patient with sporadic Parkinson’s disease[J]. Stem Cell Research, 2024, 77: 103416.
[49]	NUTTLE X, BURT N D, CURRALL B, et al. Parallelized engineering of mutational models using piggyBac transposon delivery of CRISPR libraries[J]. Cell Reports Methods, 2024, 4(1): 100672.
[50]	朱思颖, 杨洋, 李鹏东, 等. 微核糖核酸簇miR-290-295促进体细胞重编程[J]. 中国生物工程杂志, 2023, 43(4): 1-9.
	ZHU S Y, YANG Y, LI P D, et al. microRNA cluster 290-295 enhances somatic cell reprogramming[J]. China Biotechnology, 2023, 43(4): 1-9.
[51]	KIM D, KIM C H, MOON J I, et al. Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins[J]. Cell Stem Cell, 2009, 4(6): 472-476.
[52]	ABUJAROUR R, DINELLA J, PRIBADI M, et al. A chemical approach facilitates CRISPRa-only human iPSC generation and minimizes the number of targeted loci required[J]. Future Science OA, 2024, 10(1): FSO964.
[53]	OKITA K, ICHISAKA T, YAMANAKA S. Generation of germline-competent induced pluripotent stem cells[J]. Nature, 2007, 448(7151): 313-317.
[54]	HUANGFU D W, OSAFUNE K, MAEHR R, et al. Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2[J]. Nature Biotechnology, 2008, 26(11): 1269-1275.
[55]	KIM J B, GREBER B, ARAÚZO-BRAVO M J, et al. Direct reprogramming of human neural stem cells by OCT4[J]. Nature, 2009, 461(7264): 649-653.
[56]	杜旭光. 猪诱导多能性干细胞的多能性维持与体内发育阻滞机制研究[D]. 北京: 中国农业大学, 2015.
	DU X G. Study on the pluripotency maintenance of pig iPS cells and hurdles of their in vivo development[D]. Beijing: China Agricultural University, 2015.
[57]	WANG Q, ALLEN B N, BOHRER L R, et al. Conditional immortalization using SV40 large T antigen and its effects on induced pluripotent stem cell differentiation toward retinal progenitor cells[J]. Stem Cells and Development, 2025, 34(1/2): 26-34.
[58]	AUER M, KLAMPFL H S, SCHLENKE P, et al. Generation of three human erythroblast-derived iPSC lines (UBTi002-A, UBTi002-B, and UBTi002-C)[J]. Stem Cell Research, 2025, 87: 103767.
[59]	YAZAWA T, IMAMICHI Y, KITANO T, et al. Expression of Chrna9 is regulated by Tbx3 in undifferentiated pluripotent stem cells[J]. Scientific Reports, 2023, 13: 1611.
[60]	RONIMA K R, DEY C, THUMMER R P. An insight into the role of GLIS1 in embryonic development, iPSC generation, and cancer[J]. Advances in Experimental Medicine and Biology, 2024, 1470: 97-113.
[61]	YANG X C, WU X L, LI W H, et al. OCT6 inhibits differentiation of porcine-induced pluripotent stem cells through MAPK and PI3K signaling regulation[J]. Zoological Research, 2022, 43(6): 911-922.
[62]	YANG F, REN Y H, LI H, et al. ESRRB plays a crucial role in the promotion of porcine cell reprograming[J]. Journal of cellular physiology, 2018, 233(2): 1601-1611.
[63]	OGOREVC J, OREHEK S, DOVČ P. Cellular reprogramming in farm animals: An overview of iPSC generation in the mammalian farm animal species[J]. Journal of Animal Science and Biotechnology, 2016, 7: 10.
[64]	ZHOU M, ZHANG M L, GUO T X, et al. Species origin of exogenous transcription factors affects the activation of endogenous pluripotency markers and signaling pathways of porcine induced pluripotent stem cells[J]. Frontiers in Cell and Developmental Biology, 2023, 11: 1196273.
[65]	SUN Y C, HU X L, HUANG X W, et al. Hypertranscription of rDNA responsible for nucleolar remodelling is a doorman for acquiring pluripotency[J]. Cell Proliferation, 2025, 58(10): e70052.
[66]	WANG S, XIA P Y, YE B Q, et al. Transient activation of autophagy via Sox2-mediated suppression of mTOR is an important early step in reprogramming to pluripotency[J]. Cell Stem Cell, 2013, 13(5): 617-625.
[67]	MACCARTHY C M, WU G M, MALIK V, et al. Highly cooperative chimeric super-SOX induces naive pluripotency across species[J]. Cell Stem Cell, 2024, 31(1): 127-147.e9.
[68]	CHEN K S, LONG Q, XING G S, et al. Heterochromatin loosening by the Oct4 linker region facilitates Klf4 binding and iPSC reprogramming[J]. The EMBO Journal, 2019, 39(1): EMBJ201899165.
[69]	LI S S, CHEN G Y, HUANG X, et al. C-Myc alone is enough to reprogram fibroblasts into functional macrophages[J]. Journal of Hematology & Oncology, 2024, 17(1): 83.
[70]	WU X-L, ZHU Z-S, et al. LIN28A inhibits DUSP family phosphatases and activates MAPK signaling pathway to maintain pluripotency in porcine induced pluripotent stem cells[J]. Zoological Research, 2021, 42(3): 377-388.
[71]	ZHANG P F, CHEN X X, ZHENG Y, et al. Long-term propagation of porcine undifferentiated spermatogonia[J]. Stem Cells And Development, 2017, 26(15): 1121-1131.
[72]	曾虹, 曾睿琳, 付伟, 等. 牛诱导多能干细胞的建立及应用研究进展[J]. 生物技术通报, 2023, 39(5): 130-141.
	ZENG H, ZENG R L, FU W, et al. Research progress in the application and establishment of bovine induced pluripotent stem cells[J]. Biotechnology Bulletin, 2023, 39(5): 130-141.
[73]	王佳美, 黄永震, 高晨, 等. 多能性干细胞概述及其在家畜上的研究进展[J]. 畜牧兽医学报, 2025, 56(4): 1473-1483.
	WANG J M, HUANG Y Z, GAO C, et al. Research progress and overview on pluripotent stem cells in livestock[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1473-1483.
[74]	刘琴. 优化的小鼠STO细胞系重编程多潜能干细胞的建立[D]. 杨凌: 西北农林科技大学, 2012.
	LIU Q. Establishing of the optimization of mouse STO cell lines reprogram to pluripotent stem cells[D]. Yangling: Northwest A & F University, 2012.
[75]	YU G, OKAWA H, OKITA K, et al. Gingival fibroblasts as autologous feeders for induced pluripotent stem cells[J]. Journal of Dental Research, 2016, 95(1): 110-118.
[76]	HAVASI P, NABIONI M, SOLEIMANI M, et al. Mesenchymal stem cells as an appropriate feeder layer for prolonged in vitro culture of human induced pluripotent stem cells[J]. Molecular Biology Reports, 2013, 40(4): 3023-3031.
[77]	JIANG Y, CAI N N, AN X L, et al. Naïve-like conversion of bovine induced pluripotent stem cells from Sertoli cells[J]. Theriogenology, 2023, 196: 68-78.
[78]	YANG J, RYAN D J, WANG W, et al. Establishment of mouse expanded potential stem cells[J]. Nature, 2017, 550(7676): 393-397.
[79]	HUANG B, LI T, ALONSO-GONZALEZ L, et al. A virus-free poly-promoter vector induces pluripotency in quiescent bovine cells under chemically defined conditions of dual kinase inhibition[J]. PLoS One, 2011, 6(9): e24501.
[80]	HOU P P, LI Y Q, ZHANG X, et al. Pluripotent stem cells induced from mouse somatic cells by small-molecule compounds[J]. Science, 2013, 341(6146): 651-654.
[81]	RODRÍGUEZ A, ALLEGRUCCI C, ALBERIO R. Modulation of pluripotency in the porcine embryo and iPS cells[J]. PLoS One, 2012, 7(11): e49079.
[82]	SAITO S, YOKOYAMA K, TAMAGAWA T, et al. Derivation and induction of the differentiation of animal ES cells as well as human pluripotent stem cells derived from fetal membrane[J]. Human Cell, 2005, 18(3): 135-141.
[83]	BALBASI E, GUVEN G, TERZI CIZMECIOGLU N. Mouse embryonic stem cell culture in serum-containing or 2i conditions[J]. Methods in Molecular Biology, 2022, 2520: 275-294.
[84]	PETKOV S, HYTTEL P, NIEMANN H. The small molecule inhibitors PD0325091 and CHIR99021 reduce expression of pluripotency-related genes in putative porcine induced pluripotent stem cells[J]. Cellular Reprogramming, 2014, 16(4): 235-240.
[85]	ZHOU H Y, LI W L, ZHU S Y, et al. Conversion of mouse epiblast stem cells to an earlier pluripotency state by small molecules[J]. Journal of Biological Chemistry, 2010, 285(39): 29676-29680.
[86]	HU W T, YAN Q Y, FANG Y, et al. Transient folate deprivation in combination with small-molecule compounds facilitates the generation of somatic cell-derived pluripotent stem cells in mice[J]. Journal of Huazhong University of Science And Technology [medical sciences], 2014, 34(2): 151-156.
[87]	赵文轩, 高雪, 余大为, 等. 蒙山牛诱导多能干细胞的建立[J]. 畜牧兽医学报, 2025, 56(4): 1731-1743.
	ZHAO W X, GAO X, YU D W, et al. Establishment and pluripotency analysis of induced pluripotent stem cells from Mengshan cattle[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1731-1743.
[88]	ZHI M L, ZHANG J Y, TANG Q Z, et al. Generation and characterization of stable pig pregastrulation epiblast stem cell lines[J]. Cell Research, 2021, 32(4): 383-400.
[89]	ZHU Q Q, WANG F C, GAO D F, et al. Generation of stable integration-free pig induced pluripotent stem cells under chemically defined culture condition[J]. Cell Proliferation, 2023, 56(11): e13487.
[90]	YANG Y, LIU B, XU J, et al. Derivation of pluripotent stem cells with in vivo embryonic and extraembryonic potency[J]. Cell, 2017, 169(2): 243-257.e25.
[91]	XU J J, YU L Q, GUO J X, et al. Generation of pig induced pluripotent stem cells using an extended pluripotent stem cell culture system[J]. Stem Cell Research & therapy, 2019, 10(1): 193.
[92]	XU W Q, HAO R F, WANG J, et al. Methanol fixed feeder layers altered the pluripotency and metabolism of bovine pluripotent stem cells[J]. Scientific Reports, 2022, 12: 9177.
[93]	SHAFA M, SJONNESEN K, YAMASHITA A, et al. Expansion and long-term maintenance of induced pluripotent stem cells in stirred suspension bioreactors[J]. Journal of Tissue Engineering and Regenerative Medicine, 2012, 6(6): 462-472.
[94]	BURRELL K, DARDARI R, GOLDSMITH T, et al. Stirred suspension bioreactor culture of porcine induced pluripotent stem cells[J]. Stem Cells and Development, 2019, 28(18): 1264-1275.
[95]	SUGIMOTO K, YOSHIZAWA Y, YAMADA S, et al. Effects of hypoxia on pluripotency in murine iPS cells[J]. Microscopy Research and Technique, 2013, 76(10): 1084-1092.
[96]	LIU K, JI G Z, MAO J, et al. Generation of porcine-induced pluripotent stem cells by using OCT4 and KLF4 porcine factors[J]. Cellular Reprogramming, 2012, 14(6): 505-513.
[97]	BESSI B W, BOTIGELLI R C, PIERI N C G, et al. Cattle in vitro induced pluripotent stem cells generated and maintained in 5 or 20% oxygen and different supplementation[J]. Cells, 2021, 10(6): 1531.
[98]	SETTHAWONG P, PHAKDEEDINDAN P, TECHAKUMPHU M, et al. Molecular signature and colony morphology affect in vitro pluripotency of porcine induced pluripotent stem cells[J]. Reproduction in Domestic Animals, 2021, 56(8): 1104-1116.
[99]	VERMA R, LEE Y, SALAMONE D F. iPSC technology: An innovative tool for developing clean meat, livestock, and frozen ark[J]. Animals, 2022, 12(22): 3187.
[100]	YUE W, ZHANG J Q, WU X L, et al. Development of porcine induced pluripotent stem cells with a CD163 reporter system[J]. Chinese Journal of Biotechnology, 2023, 39(1): 192-203.
[101]	ZHAO Y Y, WU X L, YUAN L M, et al. Construction of CD163-knockout induced pluripotent stem cells using reprogramming technology[J]. Theriogenology, 2026, 250: 117722.
[102]	BATTY L, PARK J, QIN L F, et al. Vascular endothelial cells derived from transgene-free pig induced pluripotent stem cells for vascular tissue engineering[J]. Acta Biomaterialia, 2025, 193: 171-184.
[103]	RECCHIA K, WATHIKTHINNAKON M, BRESSAN F F, et al. Generation of bovine iPSCs from fetal fibroblasts for in vitro myogenesis and cultured meat[J]. Frontiers in Nutrition, 2025, 12: 1562981.

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物种 Species	载体类型Carrier type	重编程因子 Reprogramming factors	参考文献Regerences
猪 Swine	慢病毒载体	人OSKMLN	[7]
		人OSKM、BRG1	[32]
		人OSKM、猪TBX3	[33]
	逆转录病毒载体	人OSKM	[8]
		鼠OSK、miR302a、miR302b、miR200c	[34]
		人OSKM、TERT	[35]
	转座子载体	人OSKM	[36]
		人OSKMLN、SV40T、miR302/367	[37]
		人OSKMLN、NR5A2、BAF60A、miR302/367	[38]
牛 Cattle	逆转录病毒载体	牛OSKMLN、人OSKM	[39]
		OSKMLN、KDM4A	[31]
		人OSKM	[40]
	转座子载体	牛miR302/367	[41]
		OSKM	[21]
		鼠OSKM、人OSKMLN	[42]
羊 Sheep and goat	慢病毒载体	鼠OSKM	[9]
		人OSKMLN、SV40T	[10]
		人OSKMLN、TERT、miR200c-141	[43]
	mRNA载体	OSKM	[44]

重编程因子Reprogramming factors	作用机制 Mechanism of action	参考文献References
Oct4	激活rDNA转录与核仁重塑	[65]
Sox2	重编程早期下调mTOR的表达，并启动细胞自噬；与Oct4协同作用	[66-67]
Klf4	激活抗凋亡基因Bcl-2，调控p21表达，增加体细胞存活率；参与染色质重塑	[68]
c-Myc	通过Srebp-1调控细胞代谢转换	[69]
NANOG	通过调控DNA甲基化和组蛋白修饰维持多能性基因的染色质开放状态	[10]
LIN28	抑制let-7、激活MAPK代谢通路，协同重编程因子维持细胞的未分化状态	[70]
Oct6	Oct4的功能替代或辅助因子，调节MAPK及PI3K信号通路	[59]
SV40T	通过抑制p53通路延长细胞寿命，提高重编程效率	[57]
L-Myc	c-Myc的安全替代因子，降低重编程致瘤性	[60]
Tbx3	激活H3K4me3甲基转移酶	[59]
Glis1	诱导多水平的表观遗传和代谢重塑，介导“表观组-代谢组-表观组”的跨界级联反应	[71]

培养基Medium	配方 Formula	特殊成分 Special ingredients
DMEM/F12	1∶1体积比的DMEM及Ham's F12	Ham's F12含血清白蛋白、转铁蛋白、胰岛素及种类、浓度精确的氨基酸及微量元素
KnockOut™DMEM（KO-DMEM）	谷氨酰胺替代物、高葡萄糖DMEM（4.5 g/L）、非必需氨基酸NEAA、敲除血清替代物、FGF2、LIF、β-巯基乙醇	KSR成分相对明确、无动物源成分，能够代替血清的功能，且排除了有害成分和过量的分化诱导因子
mTeSR™ 1	DMEM/F12，TGF-β、白蛋白、脂类	—
Essential 8™	DMEM/F12、L-抗坏血酸、磷酸镁、硒钠、FGF2、胰岛素、NaHCO₃、转铁蛋白	—
N2B27	1∶1体积比的DMEM/F12和Neurobasal，B27 1%、N2 0.5%、GlutaMAX 0.5%、β-巯基乙醇55 μmol/L、青霉素-链霉素100 U/mL	Neurobasal是专为神经细胞培养设计的基础培养基。可维持神经细胞的长期生长和正常表型及高纯度的神经元培养

类型 Type	小分子化合物Small molecule compounds	作用机制 Mechanism of action	物种依赖Spectes dependence
类型 Type	小分子化合物Small molecule compounds	作用机制 Mechanism of action	猪	牛	羊
表观遗传调节剂Epigenetic modulator	VPA	组蛋白去乙酰化酶抑制剂	√	√	—
表观遗传调节剂Epigenetic modulator	丁酸钠	组蛋白去乙酰化酶抑制剂	√	—	—
信号通路激活或抑制剂 Activators and inhibitors of signaling pathways	CHIR99021	Wnt/β-catenin通路激动剂，GSK3i通路抑制剂	√	√	√
	PD0325901	MAPK/ERK通路抑制剂	√	√	√
	A8301	TGF-β/Smad通路抑制剂	√	—	—
	Y-27632	ROCK通路抑制剂	√	√	√
	WH-4-023	SRC抑制剂	—	—	—
	IWR-1-endo	Wnt/β-catenin通路抑制剂	—	—	√
	盐酸米诺环素	NF-κB通路抑制剂	—	—	—
	(S)-(+)-马来酸二甲苯茚酸酯	VIP通路抑制剂	—	—	—
细胞因子或细胞代谢调节剂Cytokine or cellular metabolic modulator	LIF	激活STAT3信号通路并抑制糖原合成酶激酶3 MAP激酶/ERK激酶信号通路	○	○	√